Design and Methods

To determine whether mutations of truncated PDGFR-alpha functional domaine reduce the activation of the PDGFRÂ°Valpha in spontaneously hypertensive rats (SHR), we constructed truncated PDGFR-alpha by gene recombination (Ad5CMV-PDGFR-alpha). Immunoblotting was used to assess tyrosine phosphorylation of PDGFR-alpha immunoprecipitated from cultured rat aortic VSMC, which were transfected with either empty vector for control or transfected with PDGFR-alpha encoding adenoviruses. Incorporation of 3H-thymidine and 3H-Leucine was measured, and intracellular free calcium in VSMC was obtained using fura-2 spectrofluorometry. Vascular reactivity of aortic rings was studied using an isometric force transducer.